Nanobody Production

NanobodyOrderForms 2020.pdfNanobodyOrderForm S2020.pdfNanobodyOrderForm AW 2.pdf

Nanobodies®: The "hot" new research tool.

We will make Nanobodies® for you.

About Nanobody Production

Single-domain antibodies (nanobodies) are becoming increasingly popular as reagents for biomedical research for  therapeutic use, and as reagents for diagnostics. The number of publications citing antibodies is increasing each year with over 2,000 publications cited in PubMed to date. This is due, in large part, to their small size, stability, high affinity, high specificity, ease of manipulation, and ease of production.  

Through resources provided by the National Institute of General Medical Sciences, our university-based Protein Core is producing single domain antibodies in alpacas on a fee for service structure. Our facility immunizes alpacas with your protein of interest, amplifies the produced pool of nanobody cDNAs by PCR and generates a phagemid library, which is then screened by panning against your protein. Positive clones are sequenced, with nanobodies to your target confirmed by ELISA. Deliverables include the nanobodies to your protein, produced and purified from E. Coli with 1 mg of purified nanobody, the nanobody clone, and its sequence. To date we have successfully generated more than 75 nanobodies. These nanobodies are for research purposes.

In many cases our core can produce your protein for nanobody production by expressing it in E. coli, insect cells, or mammalian cells and purifying it.

The next round of nanobody production will start in early November, 2021. The cost to produce a nanobody is $9,000. To produce a nanobody to your protein we need approximately 1.5 mg of purified protein in PBS or HBS at a concentration of 1 mg/ml or higher. We typically deliver your nanobody within 4 – 6 months. Usually we identify more than one nanobody against a particular protein which can be purchased for a nominal cost. A range of derivatives and conjugates are also available. 

To request a nanobody produced to your protein please fill out and return this request form:

Nanobody Production Request Form

or contact Lou Hersh at lhersh@uky.edu or 859-536-0034.

 

 

EXAMPLES OF NANOBODY USES
Determination of a Novel Protein Conform/Structure of nano body bound to a receptor reveals a previously open conformation.
Localization of an Intracellular Compartment/Nanobody (intrabody) shows protein localization to endosomes.
Pull-Down of a target protein.

SUGGESTED READING

'Nanobodies: Next Generation of Cancer Diagnostics and Therapeutics.' Emily Y. Yang and Khalid Shah. Front. Oncol., 23 July 2020

'Neutralizing nanobodies bind SARS-CoV-2 spike RBD and block interaction with ACE2.' Huo et al. Nat. Struct. Biol. 2020 Sep;27(9):846-854.

'Immunization of Alpacas (Lama Pacos) with Protein Antigens and Production of Antigen-specific Single-Domain Antibodies.' K. Martin Chow, Sidney W. Whiteheart, Jeffrey R. Smiley, Savita Sharma, Kathy Boaz, Meggie J. Coleman, Alvina Maynard, Louis B. Hersh, Craig W. Vander Kooi. J. Vis Exp 2019 Jan 26;(14

'Nanobody approval gives domain antibodies a boost.'  Morrison C. Nat Rev Drug Discov. 2019 Jul;18 (7) : 485-487

'Exploiting Nanobodies' Singular Traits.' Ingram JR, Schmidt FI, Ploegh HL. Annu Rev Immunol. 2018 Feb 28.

'Reporter-nanobody fusions (RANbodies) as versatile, small, sensitive immunohistochemical reagents.' Yamagata M, Sanes JR. Proc Natl Acad Sci U S A. 2018 Feb 27;115(9):2126-2131.

'Editorial: Single-Domain Antibodies-Biology, Engineering and Emerging Applications.' Henry KA, MacKenzie CR. Front Immunol. 2018 Jan 23;9:41.

'Nanobodies as novel agents for cancer therapy.' Revets H, De Baetselier P, Muyldermans S. Expert Opin Biol Ther. 2005 Jan;5(1):111-24. Review

'Nanobodies and their potential applications.' Hassanzadeh-Ghassabeh G, Devoogdt N, De Pauw P, Vincke C, Muyldermans S.  Nanomedicine (Lond). 2013 Jun;8(6):1013-26.

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